Methods relating to the treatment of fibrosis

ABSTRACT

The invention provides α-helix mimetic structures of formula (I) with the definitions of A, B, D, E, G, W, R 1  and R 2  as set out in the description and a chemical library relating thereto. The compounds, pharmaceutical compositions comprising the compounds, and methods of the invention using the compounds, relate to the treatment of fibrotic diseases, such as pulmonary fibrosis.

CROSS REFERENCE TO RELATED APPLICATION(S)

The present application is a continuation of PCT International PatentApplication No. PCT/US2006/009191, filed Mar. 15, 2006, which claimspriority from U.S. Provisional Application No. 60/663,499, filed Mar.18, 2005, both of which are hereby incorporated by reference in theirentirety.

STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT

This invention was made with government support under grant R01 HL073722awarded by the National Institutes of Health. The government has certainrights in the invention.

The subject matter of this application was made with support from theUnited States Government under The National Institutes of Health, GrantNo. R01 HL073722. The U.S. Government may have certain rights.

TECHNICAL FIELD

The present invention relates generally to α-helix mimetic structuresand to a chemical library relating thereto. The invention also relatesto applications in the treatment of diseases and pharmaceuticalcompositions comprising them.

BACKGROUND OF THE INVENTION

Fibrosis can occur in the lung, liver, kidney, eye, heart, and othermajor organs of the body. Fibrosis can be due to toxic or infectiousinjury, such as cigarette smoke to the lungs or viral hepatitisinfection of the liver. The cause of some fibrotic diseases is unknown,which is the case with idiopathic pulmonary fibrosis.

Idiopathic pulmonary fibrosis (IPF) is a chronic and insidiousinflammatory disease of the lung that kills most of its victims withinfive years after diagnosis. IPF afflicts 83,000 Americans and more than31,000 new cases develop each year. It is believed that death due to IPFis greatly underreported and the considerable morbidity of IPF is notrecognized. IPF represents just one of the many fibrotic diseases thatoccurs as a result of chronic inflammation. It is estimated by theUnited States government that 45% of all deaths in the U.S. can beattributed to fibrotic disorders, and therapeutic agents are needed fortreating this condition, especially fibrotic disease of the lungs.

Pulmonary fibrosis leads to progressive scarring and lung destruction.Currently, there are five million people worldwide that are affected bypulmonary fibrosis with 50% mortality at 5 years after diagnosis(Katzenstein A and Meyers Am. J. Respir. Crit. Care Med. 1998, 157,130-1-15 and American Thoracic Society, Am. J, Respir. Care Med. 2000,161, 646, 664.). Pulmonary fibrosis is believed to be initiated byinsult to the lung parenchyma (either acute or chronic) and develop inpatients unable to effectively heal the damage (Gross T. J. N. Eng. JMed 345, 517, 2001). The fibrosis is refractory to corticosteroids andno effective therapy currently exists.

Random screening of molecules for possible activity as therapeuticagents has occurred for many years and resulted in a number of importantdrug discoveries. While advances in molecular biology and computationalchemistry have led to increased interest in what has been termed“rational drug design”, such techniques have not proven as fast orreliable as initially predicted. Thus, in recent years there has been arenewed interest and return to random drug screening. To this end,particular strides having been made in new technologies based on thedevelopment of combinatorial chemistry libraries, and the screening ofsuch libraries in search for biologically active members.

In general, combinatorial chemistry libraries are simply a collection ofmolecules. Such libraries vary by the chemical species within thelibrary, as well as the methods employed to both generate the librarymembers and identify which members interact with biological targets ofinterest. While this field is still young, methods for generating andscreening libraries have already become quite diverse and sophisticated.For example, a recent review of various combinatorial chemical librarieshas identified a number of such techniques (Dolle, J. Com. Chem., 2(3):383-433, 2000), including the use of both tagged and untagged librarymembers (Janda, Proc. Natl. Acad. Sci. USA 91:10779-10785, 1994).

Initially, combinatorial chemistry libraries were generally limited tomembers of peptide or nucleotide origin. To this end, the techniques ofHoughten et al. illustrate an example of what is termed a “dual-definediterative” method to assemble soluble combinatorial peptide librariesvia split synthesis techniques (Nature (London) 354:8486, 1991;Biotechniques 13:412-421, 1992; Bioorg. Med. Chem. Lett. 3:405-412,1993). By this technique, soluble peptide libraries containing tens ofmillions of members have been obtained. Such libraries have been shownto be effective in the identification of opioid peptides, such asmethionine- and leucine-enkephalin (Dolley and Houghten, Life Sci, 52,1509-1517, 1993), and N-acylated peptide library has been used toidentify acetalins, which are potent opioid antagonists (Dooley et al.,Proc. Natl. Acad. Sci. USA 90:10811-10815, 1993). More recently, an allD-amino acid opioid peptide library has been constructed and screenedfor analgesic activity against the mu (“μ”) opioid receptor (Dooley etal., Science 266:2019-2022, 1994).

While combinatorial libraries containing members of peptide andnucleotide origin are of significant value, there is still a need in theart for libraries containing members of different origin. For example,traditional peptide libraries to a large extent merely vary the aminoacid sequence to generate library members. While it is well recognizedthat the secondary structures of peptides are important to biologicalactivity, such peptide libraries do not impart a constrained secondarystructure to its library members.

To this end, some researchers have cyclized peptides with disulfidebridges in an attempt to provide a more constrained secondary structure(Tumelty et al., J. Chem. Soc. 1067-68, 1994; Eichler et al., PeptideRes. 7:300-306, 1994). However, such cyclized peptides are generallystill quite flexible and are poorly bioavailable, and thus have met withonly limited success.

More recently, non-peptide compounds have been developed which moreclosely mimic the secondary structure of reverse-turns found inbiologically active proteins or peptides. For example, U.S. Pat. No.5,440,013 to Kahn and published PCT WO94/03494, PCT WO01/00210A1, andPCT WO01/16135A2 to Kahn disclose conformationally constrained,non-peptidic compounds, which mimic the three-dimensional structure ofreverse-turns.

While significant advances have been made in the synthesis andidentification of conformationally constrained, reverse-turn mimetics,there remains a need in the art for small molecules, which mimic thesecondary structure of peptides. There has been also a need in the artfor libraries containing such members, as well as techniques forsynthesizing and screening the library members against targets ofinterest, particularly biological targets, to identify bioactive librarymembers. For example, U.S. Pat. No. 5,929,237 and itscontinuation-in-part U.S. Pat. No. 6,013,458 to Kahn also disclosesconformationally constrained compounds which mimic the secondarystructure of reverse-turn regions of biologically active peptides andproteins. The synthesis and identification of conformationallyconstrained α-helix mimetics and their application to diseases arediscussed in Walensky, L. D. et al Science 305, 1466, 2004; Klein, C.Br. J. Cancer. 91:1415, 2004.

Many models of pulmonary fibrosis have been developed, howeverregardless of the nature of the initial insult the stages of progressionappear to be quite similar. A generally accepted model involves damageto the endothelial and type I alveolar epithelial cells followed byinterstitial edema, deposition of fibrous materials in the alveolus inareas of loss of type I epithelial cells. It is believed that limitedproliferation of the type II cells and subsequent differentiation intotype I and Clara cells is critical to reestablishment of normal gasexchange.

Anti-inflammatory therapies (e.g. corticosteroids, interferon-γ) totreat pulmonary fibrosis have been disappointing to date due to limitedefficacy and severe adverse side effects. An important unmet need existsto identify the key molecular pathways involved in the development andprogression of pulmonary fibrotic diseases and to develop newtherapeutic agents to prevent the progression and reverse the diseaseprocess. No drugs have been approved for the treatment of any fibroticdisease in the United States. Research and development is desperatelyneeded to provide treatments to those afflicted with fibroproliferativediseases. The present invention fulfills these needs, and providesfurther related advantages by providing conformationally constrainedcompounds which mimic the secondary structure of α-helix regions ofbiologically active peptides and proteins.

SUMMARY OF THE INVENTION

In brief, the present invention is directed to conformationallyconstrained compounds, which mimic the secondary structure of α-helixregions of biologically active peptides and proteins and their use fortreating fibrosis, such as pulmonary fibrosis. This invention alsodiscloses libraries containing such compounds, as well as the synthesisand screening thereof.

The compounds of the present invention have the following generalformula (I):

Wherein A is —(C═O)—CHR³—, B is N—R₄—, D is —(C═O)—(CHR₅)— or —(C═O)—, Eis -(ZR₆)— or (C═O), G is —(XR₇)_(n)—, —(CHR₇)—(NR₈)—, —(C═O)—(XR₉)—, or—(C═O)—, W is —Y(C═O)—, —(C═O)NH—, —(SO₂)— or nothing, Y is oxygen orsulfur, X and Z is independently nitrogen or CH, n=0 or 1; and R₁, R₂,R₃, R₄, R₅, R₆, R₇, R₈, and R₉ are the same or different andindependently selected from an amino acid side chain moiety orderivative thereof, the remainder of the molecule, a linker and a solidsupport, and stereoisomers thereof.

In the embodiment wherein A is —(C═O)—CHR³—, B is —(NR₄)—, D is —(C═O)—,E is -(ZR₆)—, and G is —(C═O)—(XR₉)—, the compounds of this inventionhave the following formula (III):

Wherein W, Y and n are as defined above, Z is nitrogen or CH (when Z isCH, then X is nitrogen), and R₁, R₂, R₃, R₄, R₆, and R₉ are as definedin the following detailed description.

In the embodiment wherein A is —(C═O)—(CHR₃), B is —(CHR₄)—, D is—(C—O)—, E is -(ZR₆)—, and G is (XR₇)_(n)—, the compounds of thisinvention have the following general formula (IV):

Wherein W, Y and n are as defined above, Z is nitrogen or CH (when Z isnitrogen, then n is zero, and when Z is CH, then X is nitrogen and n isnot zero), and R₁, R₂, R₄, R₆, and R₇, are as defined in the followingdetailed description.

The present invention is also directed to libraries containing compoundsof formula (I) above, as well as methods for synthesizing such librariesand methods for screening the same to identify biologically activecompounds. Compositions containing a compound of this invention incombination with a pharmaceutically acceptable carrier or diluent arealso disclosed.

Especially, the present invention relates pharmaceutical compositionscontaining compounds of formula (I) for treating disorders includingfibrosis of the lung. It further relates to methods for treatingdisorders including fibrosis of the lung which are associated with TGF-βsignaling pathway.

The compound V (ICG-001) is useful for treating fibrosis as described inExample 1.

These and other aspects of this invention will be apparent uponreference to the attached figures and following detailed description. Tothis end, various references are set forth herein, which describe inmore detail certain procedures, compounds and/or compositions, and areincorporated by reference in their entirety.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 depicts lung sections taken from Bat-Gal transgenic mice givenintratracheal saline or bleomycin and either treated with ICG-001 (5mgs/Kg/day subcutaneously) or saline as vehicle control. The lungs weresectioned and stained with X-Gal (blue color.) FIG. 1 A) intratrachealbleo+saline; FIG. 1 B) intracheal bleo+ICG-001; FIG. 1 C) saline+saline.

FIG. 2 depicts lung sections taken from C57/B16 mice treated withintratracheal bleomeycin (lower left) or saline (upper left) for 5 daysand stained with trichrome (red color) to stain collagen.

FIG. 3 shows RT-PCR data for S100A4 and collagen 1A2, which areincreased in the bleomycin treated mice (treated with saline control).Message is reduced essentially to negative control (i.e. saline/salinemice) levels by ICG-001 treatment (5 mgs/Kg/day s.c.)

FIG. 4 shows the results of IPF patient fibroblasts after culture inRPMI 1640+10% FBS for 2 days and treatment with ICG-001. Western blotsfor S100A4 (also known as FSP-1 or fibroblast specific protein-1) andE-Cadherin were performed on whole cell lysates. ICG-001 decreasedS100A4 expression and increased E-Cadherin expression.

DETAILED DESCRIPTION OF THE INVENTION

Transforming growth factor β (TGFβ), a key mediator in the developmentof fibrosis, is important in cell proliferation and differentiation,apoptosis, and deposition of extracellular matrix (ECM). TGFβ signalingactivates both the Smad and AP-1 transcription pathways. TGFβ in theairways of patients with pulmonary fibrosis (PF) may function initiallyas a “healing molecule” involved in the diminution of initial airwayinflammation and in tissue repair. However, with continued inflammatoryresponse such as may occur in PF, the balance may be shifted, toexcessive ECM deposition and development of airway fibrosis.

Fibroproliferative diseases are generally caused by the activation ofresident stellate cells which are found in most organs. This activationof stellate cells leads to their conversion to myofibroblasts whichdisplay characteristics of muscle and non-muscle cells. Activatedstellate cells initiate inflammatory signals, principally mediatedthrough TGF-β. Inflammatory cytokines and mediators in addition toTGF-β, lead to proliferation of myofibroblasts. Stellate-derivedmyofibroblasts proliferate and replace healthy, functional organ cellswith extra-cellular matrix that exhibit muscle and connective tissuetraits. Ultimately, organ failure results when the nonfunctionalfibrotic honeycomb matrix replaces a critical number of healthy cells.

The initial cause of fibrosis is believed to be the result of injury orinsult to organ tissues. This cellular injury to organ tissues can oftenbe traced to toxic or infectious agents. Pulmonary fibrosis, orinterstitial lung disease, is often the result of smoking, chronicasthma, chronic obstructive pulmonary disease (COPD) or pneumonia.

Pulmonary fibrosis destroys the lung's ability to transport oxygen andother gases into or out of the blood. This disease modifies the delicateand elastic tissues of the lung, changing these tissues into thicker,stiff fibrous tissue. This change or replacement of the original tissueis similar to the permanent scarring that can occur to other damagedtissues. Scarring of the lung reduces the lung's ability to allow gasesto pass into or out of the blood (i.e. oxygen, carbon dioxide).Gradually, the air sacs of the lungs become replaced by fibrotic tissue.When the scar forms, the tissue becomes thicker causing an irreversibleloss of the tissue's ability to transfer oxygen into the bloodstream.Symptoms include shortness of breath, particularly with exertion;chronic dry, hacking cough; fatigue and weakness; discomfort in thechest; loss of appetite; and rapid weight loss.

Several causes of pulmonary fibrosis are known and they includeoccupational and environmental exposures. Many jobs, particularly thosethat involve mining or that expose workers to asbestos or metal dusts,can cause pulmonary fibrosis. Workers doing these kinds of jobs mayinhale small particles (like silica dusts or asbestos fibers) that candamage the lungs, especially the small airways and air sacs, and causethe scarring associated with fibrosis. Agricultural workers also can beaffected. Some organic substances, such as moldy hay, cause an allergicreaction in the lung. This reaction is called Farmer's Lung and cancause pulmonary fibrosis. Other fumes found on farms are directly toxicto the lungs.

Another cause is Sarcoidosis, a disease characterized by the formationof granulomas (areas of inflammatory cells), which can attack any areaof the body but most frequently affects the lungs. Certain medicines mayhave the undesirable side effect of causing pulmonary fibrosis, as canradiation, such as treatment for breast cancer. Connective tissue orcollagen diseases such as rheumatoid arthritis and systemic sclerosisare also associated with pulmonary fibrosis. Although genetic andfamilial factors may be involved, this cause is not as common as theother causes discussed above. In Chronic Obstructive Pulmonary Disease(COPD), connective tissue proliferation and fibrosis can characterizesevere COPD. COPD can develop as a result of smoking or chronic asthma.

When all known causes of interstitial lung disease have been ruled out,the condition is called “idiopathic” (of unknown origin) pulmonaryfibrosis (IPF). Over 83,000 Americans are living with IPF, and more than31,000 new cases develop each year. This debilitating condition involvesscarring of the lungs. The lungs' air sacs develop scar, or fibrotictissue, which gradually interferes with the body's ability to transferthe oxygen into the bloodstream, preventing vital organs and tissue fromobtaining enough oxygen to function normally.

There are several theories as to what may cause IPF, including viralillness and allergic or environmental exposure (including tobaccosmoke). These theories are still being researched. Bacteria and othermicroorganisms are not thought to be the cause of IPF. There is also afamilial form of the disease, known as familial idiopathic pulmonaryfibrosis. Additional research is being done to determine whether thereis a genetic tendency to develop the disease, as well as to determineother causes of IPF.

Patients with IPF suffer similar symptoms to those with pulmonaryfibrosis when their lungs lose the ability to transfer oxygen into thebloodstream. The symptoms include shortness of breath, particularlyduring or after physical activity; spasmodic, dry cough; gradual,unintended weight loss; fatigue and weakness; chest discomfort;clubbing, or enlargement of the ends of the fingers (or sometimes thetoes) due to a buildup of tissue. These symptoms can greatly reduce IPFpatients' quality of life. Pulmonary rehabilitation, and oxygen therapycan reduce the lifestyle-altering effects of IPF, but do not provide acure.

Other mammalian fibrotic diseases that are amenable to treatmentaccording to the invention include kidney disease, polycystic kidneydisease, renal fibrotic disease, glomerular nephritis, liver cirrhosis,nephritis associated with systemic lupus, peritoneal fibrosis, liverfibrosis, polycystic ovarian syndrome, myocardial fibrosis, Grave'sopthalmopathy, glaucoma, scarring, skin lesions, diabetic retinopathy,scleroderma, and Alzheimer's disease.

In order to develop a treatment for fibrotic disease, it is important tofocus on the common pathway to the ultimate pathology that is shared bythe disease states, regardless of cause or of tissue in which it ismanifested. β-catenin plays a role in the development of fibrosis, andcompounds that modulate this pathway are useful for treating fibrosis.

Wnt signaling plays an essential role in both the development andmaintenance of multiple organ systems including the brain, intestines,skin and lung. A number of Wnt genes including Wnt2, Wnt5a, Wnt7b, Wnt11and Wnt13 are expressed both in the developing and adult lung (MorriseyE. 2003, Am. J. Pathology, 162, 1393-7). In both epithelial (type 2pneumocytes) and mesenchymal (myofibroblasts) cells, accumulation ofnuclear β-catenin, a hallmark of activated Wnt signaling has beenobserved (Chilosi et al 2003, Am J. Pathology 162, 1495-1502).Importantly, increased proliferation of type 2 cells in IPF has beenobserved (Kawanami O et al. Lab Invest 1982, 46, 39-53 and Kasper M etal. Histol. Histopathol 1996, 11, 463-83). Furthermore, activation ofWnt signaling in the adjacent mesenchyme may further prevent the properdifferentiation of the alveolar epithelium.

The well established bleomycin induced model of pulmonary fibrosis intransgenic Bat-Gal mice was used herein to demonstrate that aberrantactivation of Wnt signaling in the lungs is induced after insult.Furthermore, utilizing a specific inhibitor of Wnt/β-catenin/CBP driventranscription (ICG-001, 5 mg/Kg/day s.c.) Wnt/β-catenin was inhibitedby >95% as judged by β-galactdsidase activity. ICG-001 is among thestructures described in detail below.

The Wnt/(catenin pathway initiates a signaling cascade critical innormal development of many organ systems including the lung (Morrisey E2003 Am J Pathology, 162, 1393-7). The hallmark of this pathway is thatit activates the transcriptional role of the multifunctional proteinβ-catenin. Canonical Wnt signaling inactivates GSK-3β, preventingβ-catenin phosphorylation. This leads to accumulation of β-catenin inthe cytoplasm and subsequent translocation to the nucleus (Behrens J,2000, Ann. NY Acad. Sci. 910, 21-33.). A key step in the activation oftarget genes is the formation of a complex between β-catenin and membersof the T-cell factor (TCF)/lymphoid enhancer factor (LEF-1) family oftranscription factors. To generate a transcriptionally active complex,β-catenin recruits the transcriptional coactivators, Creb-BindingProtein (CBP) or its closely related homolog, p300 as well as othercomponents of the basal transcription machinery.

Previously, aberrant Wnt/β-catenin signaling has been demonstrated inlung samples from patients with idiopathic pulmonary fibrosis (IPF)(Chilosi M. et al 2003 Am. J. Pathol. 162, 1495-1502) with increasednuclear β-catenin immunoreactivity and increased expression of twoTCF/β-catenin regulated genes i.e. cyclin D1 and matrilysin (MMP7). Animportant role for increased MMP7 activity in pulmonary fibrotic diseaseis expected as MMP7 (−/−) mice are protected from bleomycin-inducedpulmonary fibrosis (Zuo F et al PNAS 99, 6292, 2002).

ICG-001, a small molecule (FW 548) that selectively inhibitsTCF/O-catenin transcription in a CBP-dependent fashion, was recentlyidentified (Emami et al. Proc. Natl. Acad. Sci. USA 2004, 101, 12682-7and McMillan and Kahn Drug Discovery Today 2005, 10, 1467-74). ICG-001selectively blocks the β-catenin/CBP interaction without interferingwith the highly homologous β-catenin/p300 interaction. Using a wellestablished murine model of pulmonary fibrosis in transgenic Bat-Galmice, we now demonstrate that ICG-001 (5 mg/Kg/day) blocks >95% ofbleomycin-induced TCF/β-catenin transcription. Furthermore, ICG-011 atthis dose not only halts but reverses disease progression, as judged byreduced mortality, histopathology and endogenous gene expression. Giventhe fact that currently no effective treatments for pulmonary fibroticdisease exist) inhibition of Wnt/(catenin/CBP dependent transcriptionaccording to the invention appears to offer a novel therapeutic approachand provides industrial applicability.

Canonical Wnt signaling has been shown to promote self-renewal in avariety of tissue stem cells, including neuronal stem cells andhematopoeitic stem cells. However, activation of the canonical Wntpathway can promote or inhibit differentiation depending on theexperimental circumstances.

The present invention therefore is directed to conformationallyconstrained compounds which mimic the secondary structure of α-helixregions of biological peptide and proteins (also referred to herein as“α-helix mimetics” and chemical libraries relating thereto. Suchcompounds find use in treating fibrosis, including pulmonary fibrosis.

The α-helix mimetic structures of the present invention are useful asbioactive agents, including (but not limited to) use as diagnostic,prophylactic and/or therapeutic agents. The α-helix mimetic structurelibraries of this invention are useful in the identification of suchbioactive agents. In the practice of the present invention, thelibraries may contain from tens to hundreds to thousands (or greater) ofindividual α-helix structures (also referred to herein as “members”).

In one aspect of the present invention, a α-helix mimetic structure isdisclosed having the following formula (a):

wherein A is —(C═O)—(CHR₃)—, B is —N—R₄—, D is —(CHR₅)— or —(C═O)—, E is-(ZR₆)— or —(C═O)—, G is —(XR₇)_(n)—, —(CHR₇)—(NR₈)—, —(C═O)—(XR₉)—, or—(C═O)—, W is —Y(C═O)—, —(C═O)NH—, —(SO₂)— or nothing, Y is oxygen orsulfur, X and Z is independently nitrogen or CH, n=0 or 1; and R₁, R₂,R₃, R₄, R₅, R₆, R₇, R₈ and R₉, are the same or different andindependently selected from an amino acid side chain moiety orderivative thereof, the remainder of the molecule, a linker and a solidsupport, and stereoisomers thereof.

More specifically, R₁, R₂, R₃, R₄, R₅, R₆, R₇, R₈ and R₉, areindependently selected from the group consisting of aminoC₂₋₅alkyl,guanidineC₂₋₅alkyl, C₁₋₄alkylguanidinoC₂₋₅alkyl,diC₁₋₄alkylguanidino-C₂₋₅alkyl, amidinoC₂₋₅alkyl, C₁₋₄alkylamidinoC₂₋₅alkyl, diC₁₋₄alkylamidinoC₂₋₅alkyl, C₁₋₃alkoxy, Phenyl, substitutedphenyl (where the substituents are independently selected from one ormore of amino, amidino, guanidino, hydrazino, amidrazonyl,C₁₋₄alkylamino, C₁₋₄dialkylamino, halogen, perfluoro C₁₋₄alkyl,C₁₋₄alkyl, C₁₋₃alkoxy, nitro, carboxy, cyano, sulfuryl or hydroxyl),benzyl, substituted benzyl (where the substituents on the benzyl areindependently selected from one or more of amino, amidino, guanidino,hydrazino, amidrazonyl, C₁₋₄alkylamino, C₁₋₄dialkylamino, halogen,perfluoro C₁₋₄alkyl, C₁₋₃alkyl, nitro, carboxy, cyano, sulfuryl orhydroxyl), naphthyl, substituted naphthyl (where the substituents areindependently selected from one or more of amino, amidino, guanidino,hydrazino, amidrazonyl, C₁₋₄alkylamino, C₁₋₄dialkylamino, halogen,perfluoro C₁₋₄alkyl, C₁₋₄alkyl C₁₋₃alkoxy, nitro, carboxy, cyano,sulfuryl or hydroxyl), bisphenyl methyl, substituted bis-phenyl methyl(where the substituents are independently selected from one or more ofamino, amidino, guanidino, hydrazino, amidrazonyl, C₁₋₄alkylamino,C₁₋₄dialkylamino, halogen, perfluoro C₁₋₄alkyl, C₁₋₄alkyl, C₁₋₃alkoxy,nitro, carboxy, cyano, sulfuryl or hydroxyl), pyridyl, substitutedpyridyl, (where the substituents are independently selected from one ormore of amino, amidino, guanidino, hydrazino, amidrazonyl,C₁₋₄alkylamino, C₁₋₄dialkylamino, halogen, perfluoro C₁₋₄alkyl,C₁₋₄alkyl, C₁₋₃alkoxy, nitro, carboxy, cyano, sulfuryl or hydroxyl),pyridylC₁₋₄alkyl, substituted pyridylC₁₋₄-alkyl (where the pyridinesubstituents are independently selected from one or more of amino,amidino, guanidino, hydrazino, amidrazonyl, C₁₋₄alkylamino,C₁₋₄dialkylamino, halogen, perfluoro C₁₋₄alkyl, C₁₋₄alkyl, C₁₋₃alkoxy,nitro, carboxy, cyano, sulfuryl or hydroxyl), pyrimidylC₁₋₄alkyl,substituted pyrimidylC₁₋₄alkyl (where the pyrimidine substituents areindependently selected from one or more of amino, amidino, guanidino,hydrazino, amidrazonyl, C₁₋₄alkylamino, C₁₋₄dialkylamino, halogen,perfluoro C₁₋₄alkyl, C₁₋₄alkyl, C₁₋₃alkoxy, nitro, carboxy, cyano,sulfuryl or hydroxyl), triazin-2-yl-C₁₋₄alkyl, substitutedtriazin-2-yl-C₁₋₄alkyl (where the triazine substituents areindependently selected from one or more of amino, amidino, guanidino,hydrazino, amidrazonyl, C₁₋₄alkylamino, C₁₋₄dialkylamino, halogen,perfluoro C₁₋₄alkyl, C₁₋₄alkyl, C₁₋₃alkoxy, nitro, carboxy, cyano,sulfuryl or hydroxyl), imidazoC₁₋₄alkyl, substituted imidazol C₁₋₄alkyl(where the imidazole substituents are independently selected from one ormore of amino, amidino, guanidino, hydrazino, amidrazonyl,C₁₋₄alkylamino, C₁₋₄dialkylamino, halogen, perfluoro C₁₋₄alkyl,C₁₋₄alkyl, C₁₋₃alkoxy, nitro, carboxy, cyano, sulfuryl or hydroxyl),imidazolinylCalkyl, N-amidinopiperazinyl-N—C₀₋₄alkyl, hydroxyC₂₋₅alkyl,C₁₋₅alkylaminoC₂₋₅alkyl, hydroxyC₂₋₅alkyl, C₁₋₅alkylaminoC₂₋₅alkyl,C₁₋₅dialkylaminoC₂₋₅alkyl, N-amidinopiperidinylC₁₋₄alkyl and4-aminocyclohexylC₀₋₂alkyl.

In one embodiment, R₁, R₂, R₆ of E, and R₇, R₈ and R₉ of G are the sameor different and represent the remainder of the compound, and R₃ or A,R₄ of B or R₅ of D is selected from an amino acid side chain moiety orderivative thereof. As used herein, the term “remainder of the compound”means any moiety, agent, compound, support, molecule, linker, aminoacid, peptide or protein covalently attached to the α-helix mimeticstructure at R₁, R₂, R₅, R₆, R₇, R₈ and/or R₉ positions. This term alsoincludes amino acid side chain moieties and derivatives thereof.

As used herein, the term “amino acid side chain moiety” represents anyamino acid side chain moiety present in naturally occurring proteinsincluding (but not limited to) the naturally occurring amino acid sidechain moieties identified in Table 1. Other naturally occurring aminoacid side chain moieties of this invention include (but are not limitedto) the side chain moieties of 3,5-dibromotyrosine, 3,5-diiodotyrosine,hydroxylysine, γ-carboxyglutamate, phosphotyrosine and phosphoserine. Inaddition, glycosylated amino acid side chains may also be used in thepractice of this invention, including (but not limited to) glycosylatedthreonine, serine and asparagine.

TABLE 1 Amino Acid Side Chain Moieties Amino Acid Side Chain MoietyAmino Acid —H Glycine —CH₃ Alanine —CH(CH₃)₂ Valine —CH₂CH(CH₃)₂ Leucine—CH(CH₃)CH₂CH₃ Isoleucine —(CH₂)₄NH₃ ⁺ Lysine —(CH₂)₃NHC(NH₂)NH₂ ⁺Arginine Histidine —CH₂COO⁻ Aspartic acid —CH₂CH₂COO⁻ Glutamic acid—CH₂CONH₂ Asparagine —CH₂CH₂CONH₂ Glutamine Phenylalanine TyrosineTryptophan —CH₂SH Cysteine —CH₂CH₂SCH₃ Methionine —CH₂OH Serine—CH(OH)CH₃ Threonine Proline Hydroxyproline

In addition to naturally occurring amino acid side chain moieties, theamino acid side chain moieties of the present invention also includevarious derivatives thereof. As used herein, a “derivative” of an aminoacid side chain moiety includes modifications and/or variations tonaturally occurring amino acid side chain moieties. For example, theamino acid side chain moieties of alanine, valine, leucine, isoleucineand pheylalanine may generally be classified as lower chain alkyl, aryl,or arylalkyl moieties. Derivatives of amino acid side chain moietiesinclude other straight chain or branched, cyclic or noncyclic,substitutes or unsubstituted, saturated or unsaturated lower chainalkyl, aryl or arylalkyl moieties.

As used herein, “lower chain alkyl moieties” contain from 1-12 carbonatoms, “lower chain aryl moieties” contain from 6-12 carbon atoms and“lower chain aralkyl moieties” contain from 7-12 carbon atoms. Thus, inone embodiment, the amino acid side chain derivative is selected from aC₁₋₁₂ alkyl a C₆₋₁₂ aryl and a C₇₋₁₂ arylalkyl, and in a more preferredembodiment, from a C₁₋₇ alkyl, a C₆₋₁₀ aryl and a C₇₋₁₁ arylalkyl.

Amino side chain derivatives of this invention further includesubstituted derivatives of lower chain alkyl, aryl, and arylalkylmoieties, wherein the substituents is selected from (but are not limitedto) one or more of the following chemical moieties: —OH, —OR, —COOH,—COOR, —CON—H₂, —NH₂, —NHR, —NRR, —SH, —SR, —SO₂R, —SO₂H, —SOR andhalogen (including F, Cl, Br and I), wherein each occurrence of R isindependently selected from straight chain or branched, cyclic ornoncyclic, substituted or unsubstituted, saturated or unsaturated lowerchain alkyl, aryl, and aralkyl moieties. Moreover, cyclic lower chainalkyl, aryl and arylalkyl moieties of this invention includenaphthalene, as well as heterocyclic compounds such as thiophene,pyrrole, furan, imidazole, oxazole, thiazole, pyrazole, 3-pyrroline,pyrrolidine, pyridine, pyrimidine, purine, quinoline, isoquinoline andcarbazole. Amino acid side chain derivatives further include heteroalkylderivatives of the alkyl portion of the lower chain alkyl and aralkylmoieties, including (but not limited to) alkyl and aralkyl phosphonatesand silanes.

Representative R₁, R₂, R₅, R₆, R₇, R₈ and R₉ moieties specificallyinclude (but are not limited to) —OH, —OR, —COR, —COOR, —CONH₂, —CONR,—CONRR, —NH₂, —NHR, —NRR, —SO₂R and —COSR, wherein each occurrence of Ris as defined above.

In a further embodiment, and in addition to being an amino acid sidechain moiety or derivative thereof (or the remainder of the compound inthe case of R₁, R₂, R₅, R₆, R₇, R₈ and R₉), R₁, R₂, R₅, R₆, R₇, R₈ or R₉may be a linker facilitating the linkage of the compound to anothermoiety or compound. For example, the compounds of this invention may belinked to one or more known compounds, such as biotin, for use indiagnostic or screening assay. Furthermore, R₁, R₂, R₅, R₆, R₇, R₈ or R₉may be a linker joining the compound to a solid support (such as asupport used in solid phase peptide synthesis) or alternatively, may bethe support itself. In this embodiment, linkage to another moiety orcompound, or to a solid support, is preferable at the R₁, R₂, R₇ or R₈position, and more preferably at the R₁ or R₂ position.

In the embodiment wherein A is —(C═O)—CHR₃—, B is —N—R₄, D is —(C—O)—, Eis -(ZR₆)—, G is —(C═O)—(XR₉)—, the α-helix mimetic compounds of thisinvention have the following general formula (III):

wherein R₁, R₂, R₄, R₆, R₉, W and X are as defined above, Z is nitrogenor CH (when Z is CH, then X is nitrogen). In a preferred embodiment, R₁,R₂, R₆, and R₉ represent the remainder of the compound, and R₄ isselected from an amino acid side chain moiety. In a more specificembodiment wherein A is —O—CHR₃—, B is —NR₄—, D is —(C═O)—, E is-(ZR₆)—, Gi is (XR₇)_(n)—, the α-helix mimetic compounds of thisinvention have the following formula (IV):

wherein R₁, R₂, R₄, R₆, R₇, W, X and n are as defined above, and Z isnitrogen or CH (when Z is nitrogen, then n is zero, and when Z is CH,then X is nitrogen and n is not zero). In a preferred embodiment, R₁,R₂, R₆, and R₇ represent the remainder of the compound, and R₄ isselected from an amino acid side chain moiety. In this case, R₆ or R₇may be selected from an amino acid side chain moiety when Z and X areCH, respectively.

The α-helix mimetic structures of the present invention may be preparedby utilizing appropriate starting component molecules (hereinafterreferred to as “component pieces”). Briefly, in the synthesis of α-helixmimetic structures having formula (II), first and second componentpieces are coupled to form a combined first-second intermediate, ifnecessary, third and/or fourth component pieces are coupled to form acombined third-fourth intermediate (or, if commercially available, asingle third intermediate may be used), the combined first-secondintermediate and third-fourth intermediate (or third intermediate) arethen coupled to provide a first-second-third-fourth intermediate (orfirst-second-third intermediate) which is cyclized to yield the α-helixmimetic structures of this invention. Alternatively, the α-helix mimeticstructures of formula (II) may be prepared by sequential coupling of theindividual component pieces either stepwise in solution or by solidphase synthesis as commonly practiced in solid phase peptide synthesis.

Within the context of the present invention, a “first component piece”has the following formula S1

Wherein R₂ as defined above, and R is a protective group suitable foruse in peptide synthesis. Suitable R groups include alkyl groups and, ina preferred embodiment, R is a methyl group. Such first component piecesmay be readily synthesized by reductive amination or substitutionreaction by displacement of H₂N—R₂ from CH(OR)—CHO or CH(OR)₂—CH₂-Hal(wherein Hal means a halogen atom).

A “second component piece” of this invention has the following formulaS2:

Where L₁ is carboxyl-activation group such as halogen atom, R₃, R₄ is asdefined above, and P is an amino protective group suitable for use inpeptide synthesis. Preferred protective groups include t-butyldimethylsilyl (TBDMS), t-Butyloxycarbonyl (BOC), Methylosycarbonyl(MOC), 9H-Fluorenylmethyloxycarbonyl (FMOC), and allyloxycarbonyl(Alloc). When L is —C(O)NHR, —NHR may be a carboxyl protective group.N-hydrazino amino acids can be readily prepared according to theprocedures of Vidal et al. (Tetrahedron Letters 39:8845-8848, 1998). Theconversion of these compounds to the second component pieces of thisinvention may be readily achieved by activation of the carboxylic acidgroup of the N-protected hydrazine-amino acid. Suitable activatedcarboxylic acid groups include acid halides where X is a halide such aschloride or bromide, acid anhydrides where X is an acyl group such asacetyl, reactive esters such as an N-hydroxysuccinimide esters andpentafluorophenyl esters, and other activated intermediates such as theactive intermediate formed in a coupling reaction using a carbodiimidesuch as dicyclohexylcarbodiimide (DCC).

A “third component piece” of this invention has the following formulaS3:

where G, E, and L₁ are as defined above. Suitable third component piecesare commercially available from a variety of sources or can be preparedby known methods in organic chemistry.

More specifically, the α-helix mimetic structures of this invention offormula (II) are synthesized by reacting a first component piece with asecond component piece to yield a combined first-second intermediate,followed by either reacting the combined first-second intermediate withthird component pieces sequentially to provide a combinedfirst-second-third-fourth intermediate, and the cyclizing thisintermediate to yield the α-helix mimetic structure.

The general synthesis of a α-helix having structure I′ may besynthesized by the following technique. A first component piece 1 iscoupled with a second component piece 2 by using coupling reagent suchas phosgene to yield, after N-deprotection, a combined first-secondintermediate 1-2 as illustrated below:

wherein R₁, R₂, R₄, R₇.Fmoc, Moc and X are as defined above, and Polrepresents a polymeric support.

The synthesis of representative component pieces of this invention aredescribed in Preparation Examples and Working Examples.

The α-helix mimetic structures of formula (III) and (IV) may be made bytechniques analogous to the modular component synthesis disclosed above,but with appropriate modifications to the component pieces.

As mentioned above, the reverse-turn mimetics of U.S. Pat. No. 6,013,458to Kahn, et al. are useful as bioactive agents, such as diagnostic,prophylactic, and therapeutic agents. The opiate receptor bindingactivity of representative reverse-turn mimetics is presented in Example9 of said U.S. Pat. No. 6,013,458, wherein the reverse-turn mimetics ofthis invention were found to effectively inhibit the binding of aradiolabeled enkephalin derivative to the 6 and g opiate receptors, ofwhich data demonstrates the utility of these reverse-turn mimetics asreceptor agonists and as potential analgesic agents.

The α-helix mimetic structures of the present invention will be usefulas bioactive agents, such as diagnostic, prophylactic, and therapeuticagents.

Therefore, since the compounds according to the present invention are ofα-helix mimetic structures, it may be useful for modulating a cellsignaling transcription factor related peptides in a warm-bloodedanimal, comprising administering to the animal an effective amount ofthe compound of formula (I).

A particular compound, referred to as ICG-001, is shown below ascompound V:

Further, the α-helix mimetic structures of the present invention mayalso be effective for inhibiting transcription factor/coactivator andtranscription factor corepressor interactions.

Therefore, it is an object of the present invention to provide apharmaceutical composition comprising a safe and effective amount of thecompound having general formula (VI) and pharmaceutically acceptablecarrier, which can be used for treatment of fibrotic disorders modulatedby TGF-β signaling pathway.

In another aspect of this invention, libraries containing α-helixmimetic structures of the present invention are disclosed. Onceassembled, the libraries of the present invention may be screened toidentify individual members having bioactivity. Such screening of thelibraries for bioactive members may involve; for example, evaluating thebinding activity of the members of the library or evaluating the effectthe library members have on a functional assay. Screening is normallyaccomplished by contacting the library members (or a subset of librarymembers) with a target of interest, such as, for example, an antibody,enzyme, receptor or cell line. Library members, which are capable ofinteracting with the target of interest, are referred to herein as“bioactive library members” or “bioactive mimetics”. For example, abioactive mimetic may be a library member which is capable of binding toan antibody or receptor, which is capable of inhibiting an enzyme, orwhich is capable of eliciting or antagonizing a functional responseassociated, for example, with a cell line. In other words, the screeningof the libraries of the present invention determines which librarymembers are capable of interacting with one or more biological targetsof interest. Furthermore, when interaction does occur, the bioactivemimetic (or mimetics) may then be identified from the library members.The identification of a single (or limited number) of bioactivemimetic(s) from the library yields α-helix mimetic structures which arethemselves biologically active, and thus useful as diagnostic,prophylactic or therapeutic agents, and may further be used tosignificantly advance identification of lead compounds in these fields.

In another aspect of this invention, methods for constructing thelibraries are disclosed. Traditional combinatorial chemistry techniques(see, e.g., Gallop et al., J. Med. Chem. 37:1233-1251, 1994) permit avast number of compounds to be rapidly prepared by the sequentialcombination of reagents to a basic molecular scaffold. Combinatorialtechniques have been used to construct peptide libraries derived fromthe naturally occurring amino acids. For example, by taking 20 mixturesof 20 suitably protected and different amino acids and coupling eachwith one of the 20 amino acids, a library of 400 (i.e., 20²) dipeptidesis created. Repeating the procedure seven times results in thepreparation of a peptide library comprised of about 26 billion (i.e.,20⁸) octapeptides.

Specifically, synthesis of the peptide mimetics of the library of thepresent invention may be accomplished using known peptide synthesistechniques, for example, the General Scheme of [4,4,0] α-helix MimeticLibrary as follows:

Synthesis of the peptide mimetics of the libraries of the presentinvention was accomplished using a FlexChem Reactor Block which has 96well plates by known techniques. In the above scheme ‘Pol’ represents abromoacetal resin (Advanced ChemTech) and detailed procedure isillustrated below.

Step 1

A bromoacetal resin (37 mg, 0.98 mmol/g) and a solution of R₂-amine inDMSO (1.4 mL) were placed in a Robbins block (FlexChem) having 96 wellplates. The reaction mixture was shaken at 60° C. using a rotating oven[Robbins Scientific] for 12 hours. The resin was washed with DMF, MeOH,and then DCM

Step 2

A solution of available Fmoc hydrazine Amino Acids (4 equiv.), PyBop (4equiv.), HOAt (4 equiv.), and DIEA (12 equiv.) in DMF was added to theresin. After the reaction mixture was shaken for 12 hours at roomtemperature, the resin was washed with DMF, MeOH, and them DCM.

Step 3

To the resin swollen by DMF before reaction was added 25% piperidine inDMF and the reaction mixture was shaken for 30 min at room temperature.This deprotection step was repeated again and the resin was washed withDMF, Methanol, and then DCM. A solution of hydrazine acid (4 equiv.),HOBt (4 equiv.), and DIC (4 equiv.) in DMF was added to the resin andthe reaction mixture was shaken for 12 hours at room temperature. Theresin was washed with DMF, MeOH, and then DCM.

Step 4a (Where Hydrazine Acid is MOC Carbamate)

The resin obtained in Step 3 was treated with formic acid (1.2 mL eachwell) for 18 hours at room temperature. After the resin was removed byfiltration, the filtrate was condensed under a reduced pressure usingSpeedVac [SAVANT] to give the product as oil. The product was dilutedwith 50% water/acetonitrile and then lyophilized after freezing.

Step 4b (Where Fmoc Hydrazine Acid is Used to Make Urea ThroughIsocyanate)

To the resin swollen by DMF before reaction was added 25% piperidine inDMF and the reaction mixture was shaken for 30 min at room temperature.This deprotection step was repeated again and the resin was washed withDMF, Methanol, then DCM. To the resin swollen by DCM before reaction wasadded isocyanate (5 equiv.) in DCM. After the reaction mixture wasshaken for 12 hours at room temperature the resin was washed with DMF,MeOH, then DCM. The resin was treated with formic acid (1.2 mL eachwell) for 18 hours at room temperature. After the resin was removed byfiltration, the filtrate was condensed under a reduced pressure usingSpeedVac [SAVANT] to give the product as oil. The product was dilutedwith 50% water/acetonitrile and then lyophilized after freezing.

Step 4c (Where Fmoc-Hydrazine Acid is Used to Make Urea Through ActiveCarbamate)

To the resin swollen by DMF before reaction was added 25% piperidine inDMF and the reaction mixture was shaken for 30 min at room temperature.This deprotection step was repeated again and the resin was washed withDMF, MeOH, and then DCM. To the resin swollen by DCM before reaction wasadded p-nitrophenyl chloroformate (5 equiv.) and diisopropyl ethylamine(5 equiv.) in DCM. After the reaction mixture was shaken for 12 hours atroom temperature, the resin was washed with DMF, MeOH, and then DCM. Tothe resin was added primary amines in DCM for 12 hours at roomtemperature and the resin was washed with DMF, MeOH, and then DCM. Afterreaction the resin was treated with formic acid (1.2 mL each well) for18 hours at room temperature. After the resin was removed by filtration,the filtrate was condensed under a reduced pressure using SpeedVac[SAVANT] to give the product as oil. The product was diluted with 50%water/acetonitrile and then lyophilized after freezing.

It should be understood that the examples and embodiments describedherein are for illustrative purposes only and that various modificationsor changes in light thereof will be suggested to persons skilled in theart and are to be included within the spirit and purview of thisapplication.

EXAMPLE 1 Effect of ICG-001 on Pulmonary Fibrosis

Murine models of bleomycin induced fibrosis have been developed in orderto study fibrotic disease progression. Bleomycin induced murine fibrosishas been shown to lead to aberrant alveolar epithelial repair, withincreased metaplastic alveolar cells that apparently do not properlydifferentiate to a type I phenotype (Adamson and Bowden 1979 Am JPathol. 1979 August; 96(2):531-44.). Utilizing this model, it isdemonstrated in this Example that the Wnt/β-catenin pathway plays acritical role in the development of pulmonary fibrosis and validatesthat the inhibition of this pathway with ICG-001 represents a therapyfor the treatment of pulmonary fibrotic disease.

Using this murine model of pulmonary fibrosis in transgenic Bat-Galmice, ICG-001 (5 mg/Kg/day) blocked >95% of bleomycin-inducedTCF/β-catenin transcription. Furthermore, ICG-001 at this dose not onlyhalted but reversed disease progression, as judged by reduced mortality,histopathology and endogenous gene expression.

FIG. 1 shows lung sections taken from Bat-Gal transgenic mice. Thesemice have a Beta-Galactosidase transgene driven by a TCF/Catenin drivenpromoter (i.e. a read out for activated Wnt/catenin signaling). The micewere given intratracheal saline or bleomycin and either treated withICG-001 (5 mgs/Kg/day subcutaneously) or saline as vehicle control. Themice were sacrificed and the lungs sectioned and stained with X-Gal(blue color) A) intratracheal bleo+saline. B) intracheal bleo+ICG-001 C)saline+saline.

The dose was selected because ICG-001 reduces TCF/>Catenin drivenO-Galactosidase expression >95% at 5 mgs/Kg/day.

FIG. 2 shows lung sections taken from C57/B16 mice treated withintratracheal bleomeycin (lower left) or saline (upper left) for 5 daysand stained with trichrome (red color) to stain collagen. There is anabsence of airway epithelium in lower left compared to upper left (seearrow heads) and extensive collagen deposition (lower left). On thesixth day, either saline (upper right) or ICG-001 (5 mgs/Kg/day) wasadministered for 10 days after which the mice were sacrificed andsectioned. Of interest is the upper right (saline treatment) showinglack of normal airway epithelialization, extensive collagen depositionand intra-airway hypercellularity (fibroblasts and inflammatory influx).After treatment with ICG-001, the airway looks essentially normal(compare to untreated (saline) control) (upper left), with normalcollagen levels. The mice also regained normal body weight and survived(untreated controls did not).

FIG. 3 shows RT-PCR data for S100A4 and collagen 1A2, which areincreased in the bleomycin treated mice (treated with saline control).Message is reduced essentially to negative control (i.e. saline/salinemice) levels by ICG-001 treatment (5 mgs/Kg/day s.c.)

As shown in FIG. 4, IPF patient fibroblasts were cultured inRPMI1640+10% FBS for 2 days and treated with ICG-001. Western blots forS100A4 (also know as FSP-1 or fibroblast specific protein-1) andE-Cadherin were performed on whole cell lysates. ICG-001 decreasedS100A4 expression and increased E-cadherin expression (this is also trueat the mRNA level). These data demonstrate that ICG-001 mediates amesenchymal to epithelial transition that is essential for normalhealing, re-epithelialization and ameliorization of fibrosis.

All patents, patent applications, provisional applications, andpublications referred to or cited herein are incorporated by referencein their entirety, including all figures and tables, to the extent theyare not inconsistent with the explicit teachings of this specification.

1. A method of treating a fibrotic disease, said method comprisingadministering to a mammal in need of said treatment at least onecompound having the following general formula (I) or a stereoisomerthereof:

wherein A is —(CHR₃)—(C═O)—; B is —(NR₄)—; D is —(CHR₅)— or —(C═O)—; Eis -(ZR₆)— or —(C═O)—, where Z is nitrogen or CH; G is —(XR₇)_(n)—,—(CHR₇)—(NR₈)—, —(C═O)—(XR₉)—, or —(C═O)—, where X is nitrogen or CH andn=0 or 1; W is —(Y)—(C═O)—, —(C═O)—(NR₈)—, —(SO₂)—, or nothing, where Yis oxygen or sulfur; and R₁, R₂, R₃, R₄, R₅, R₆, R₇, R₈ and R₉ are thesame or different and independently selected from an amino acid sidechain, a derivative of an amino acid side chain, a linker facilitatinglinkage of the compound to another moiety or compound, a linker joiningthe compound to a solid support, and a solid support; under conditionseffective to treat a fibrotic disease in the mammal.
 2. The methodaccording to claim 1, wherein the fibrotic disease is at least oneselected from the group consisting of radiation-induced pulmonaryfibrosis, chemotherapy-induced pulmonary fibrosis, obliterativebronchiolitis, and silicosis lesions.
 3. The method according to claim1, wherein the fibrotic disease is at least one of kidney disease,polycystic kidney disease, renal fibrotic disease, glomerular nephritis,liver cirrhosis, nephritis associated with systemic lupus, peritonealfibrosis, liver fibrosis, polycystic ovarian syndrome, myocardialfibrosis, Grave's opthalmopathy, glaucoma, scarring, skin lesions,diabetic retinopathy, scleroderma, and Alzheimer's disease.
 4. Themethod according to claim 1, wherein the fibrotic disease is at leastone selected from the group consisting of interstitial lung disease,fibrotic lung disease, and pulmonary fibrosis.
 5. The method of claim 1,wherein the compound is administered orally, transdermally,intravenously, by inhalation, or rectally.
 6. The method of claim 5,wherein the compound is administered orally.
 7. The method of claim 4,wherein the compound is administered in a form selected from the groupconsisting of capsules, tablets, powders, granules, syrups, injectablefluids, creams, ointments, hydrophilic ointments, inhalable fluids, eyedrops, and suppositories.
 8. The method according to claim 1, whereinR₁, R₂, R₃, R₄, R₅, R₆, R₇, R₈ and R₉ are independently selected fromthe group consisting of aminoC₂₋₅alkyl, guanidinoC₂₋₅alkyl,C₁₋₄alkylguanidinoC₂₋₅alkyl, diC₁₋₄alkylguanidino-C₂₋₅alkyl,amidinoC₂₋₅alkyl, C₁₋₄alkylamidinoC₂₋₅alkyl,diC₁₋₄alkylamidinoC₂₋₅alkyl, C₁₋₃alkoxy, Phenyl, substituted phenyl(where the substituents are independently selected from one or more ofamino, amidino, guanidino, hydrazino, amidrazonyl, C₁₋₄alkylamino,C₁₋₄dialkylamino, halogen, perfluoro C₁₋₄alkyl, C₁₋₄alkyl, C₁₋₃alkoxy,nitro, carboxy, cyano, sulfuryl, and hydroxyl), benzyl, substitutedbenzyl (where the substituents on the benzyl are independently selectedfrom one or more of amino, amidino, guanidino, hydrazino, amidrazonyl,C₁₋₄alkylamino, C₁₋₄-dialkylamino, halogen, perfluoro C₁₋₄alkyl,C₁₋₄alkyl, C₁₋₃alkoxy, nitro, carboxy, cyano, sulfuryl, and hydroxyl),naphthyl, substituted naphthyl (where the substituents are independentlyselected from one or more of amino, amidino, guanidino, hydrazino,amidrazonyl, C₁₋₄alkylamino, C₁₋₄dialkylamino, halogen, perfluoroC₁₋₄alkyl, C₁₋₄alkyl, C₁₋₃alkoxy, nitro, carboxy, cyano, sulfuryl, andhydroxyl), bis-phenyl methyl, substituted bis-phenyl methyl (where thesubstituents are independently selected from one or more of amino,amidino, guanidino, hydrazino, amidrazonyl, C₁₋₄alkylamino,C₁₋₄dialkylamino, halogen, perfluoro C₁₋₄alkyl, C₁₋₄alkyl, C₁₋₃alkoxy,nitro, carboxy, cyano, sulfuryl, and hydroxyl), pyridyl, substitutedpyridyl (where the substituents are independently selected from one ormore of amino, amidino, guanidino, hydrazino, amidrazonyl,C₁₋₄alkylamino, C₁₋₄dialkylamino, halogen, perfluoro C₁₋₄alkyl,C₁₋₄alkyl, C₁₋₃alkoxy, nitro, carboxy, cyano, sulfuryl, and hydroxyl),pyridylC₁₋₄alkyl, substituted pyridylC₁₋₄alkyl (where the pyridinesubstituents are independently selected from one or more of amino,amidino, guanidino, hydrazino, amidrazonyl, C₁₋₄alkylamino,C₁₋₄dialkylamino, halogen, perfluoro C₁₋₄alkyl, C₁₋₄alkyl, C₁₋₃alkoxy,nitro, carboxy, cyano, sulfuryl, and hydroxyl), pyrimidylC₁₋₄alkyl,substituted pyrimidylC₁₋₄alkyl (where the pyrimidine substituents areindependently selected from one or more of amino, amidino, guanidino,hydrazino, amidrazonyl, C₁₋₄alkylamino, C₁₋₄dialkylamino, halogen,perfluoro C₁₋₄alkyl, C₁₋₄alkyl, C₁₋₃alkoxy, nitro, carboxy, cyano,sulfuryl, and hydroxyl), triazin-2-yl-C₁₋₄alkyl, substitutedtriazin-2-yl-C₁₋₄alkyl (where the triazine substituents areindependently selected from one or more of amino, amidino, guanidino,hydrazino, amidrazonyl, C₁₋₄alkylamino, C₁₋₄dialkylamino, halogen,perfluoro C₁₋₄alkyl, C₁₋₄alkyl, C₁₋₃alkoxy, nitro, carboxy, cyano,sulfuryl, and hydroxyl), imidazoC₁₋₄alkyl, substituted imidazolC₁₋₄alkyl (where the imidazole substituents are independently selectedfrom one or more of amino, amidino, guanidino, hydrazino, amidrazonyl,C₁₋₄alkylamino, C₁₋₄dialkylamino, halogen, perfluoro C₁₋₄alkyl,C₁₋₄alkyl, C₁₋₃alkoxy, nitro, carboxy, cyano, sulfuryl, and hydroxyl),imidazolinylC₁₋₄alkyl, N-amidinopiperazinyl-N—C₀₋₄alkyl,hydroxyC₂₋₅alkyl, C₁₋₅alkylaminoC₂₋₅alkyl, hydroxyC₂₋₅alkyl,C₁₋₅alkylaminoC₂₋₅alkyl, C₁₋₅dialkylaminoC₂₋₅alkyl,N-amidinopiperidinylC₁₋₄alkyl, and 4-aminocyclohexylC₀₋₂alkyl.
 9. Themethod according to claim 1, wherein the compound has the followinggeneral formula (III):

with the proviso that when Z is CH, X is nitrogen.
 10. The methodaccording to claim 8, wherein the compound has the formula (IV):

wherein when Z is nitrogen, n is zero; and when Z is CH, X is nitrogenand n is 1, or X(_(n)) is zero.
 11. A method of treating a fibroticdisease, said method comprising administering to a mammal in need ofsaid treatment at least one compound having the following generalformula (V) or a stereoisomer thereof:

under conditions effective to treat a fibrotic disease in the mammal.